Polymerase-chain-reaction-based analysis of polymorphism in the HLA-B gene

Masayuki Yoshida, Akinori Kimura, Fujio Numano, Takehiko Sasazuki

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

The polymerase chain reaction (PCR) in combination with the sequence-specific oligonucleotide probe (SSOP) was applied to analyze the polymorphism in the axon 2 of the HLA-B gene. In this study, genomic DNAs from 85 B-lymphoblastoid cell lines homozygous for HLA and peripheral blood granulocytes of 156 Japanese individuals were investigated. Two HLA-B-specific 5′-sided primers (CG4 and CG5) and two 3′-sided primers (CG2 and CG3) were designed for specific amplification of the axon. HLA-B allelss were classified into two groups (groups I and II) according to specific amplification with two types of the 3′-sided primers. The amplified DNAs were hybridized with 23 nonradioactively labeled SSOPs. Based on the hybridization patterns with the SSOPs, 34 HLA-B specificities were divided into 26 epitope combination (EC) groups. Fifteen HLA-B specificities were classified into four EC groups and these HLA-B specificities could not be distinguished from one another in the same EC group. Another 16 HLA-B specificities corresponded one by one to 16 distinct EC groups, and two subtypes of HLA-Bw 75, B27, and Bw48 were also identified enabling the accurate typing of 22 HLA-B alleles at the DNA level. Single-strand conformation polymorphism (SSCP) of the PCR products from group I HLA-B alleles was also investigated. The HLA-B allelss showed distinct electrophoretic patterns in nondenaturing polyacrylamide gels, depending on the nucleotide sequences of the exon 2, indicating that the SSCP analysis may be an alternative, useful and practical HLA-matching system of HLA-B specificity in tissue transplantation.

Original languageEnglish
Pages (from-to)257-266
Number of pages10
JournalHuman Immunology
Volume34
Issue number4
DOIs
Publication statusPublished - Jan 1 1992

Fingerprint

HLA-B Antigens
Polymerase Chain Reaction
Genes
Epitopes
Axons
DNA
Alleles
Tissue Transplantation
Oligonucleotide Probes
Granulocytes
Exons

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Polymerase-chain-reaction-based analysis of polymorphism in the HLA-B gene. / Yoshida, Masayuki; Kimura, Akinori; Numano, Fujio; Sasazuki, Takehiko.

In: Human Immunology, Vol. 34, No. 4, 01.01.1992, p. 257-266.

Research output: Contribution to journalArticle

Yoshida, Masayuki ; Kimura, Akinori ; Numano, Fujio ; Sasazuki, Takehiko. / Polymerase-chain-reaction-based analysis of polymorphism in the HLA-B gene. In: Human Immunology. 1992 ; Vol. 34, No. 4. pp. 257-266.
@article{d94fb75737664d8bbaf6ec1335e9d271,
title = "Polymerase-chain-reaction-based analysis of polymorphism in the HLA-B gene",
abstract = "The polymerase chain reaction (PCR) in combination with the sequence-specific oligonucleotide probe (SSOP) was applied to analyze the polymorphism in the axon 2 of the HLA-B gene. In this study, genomic DNAs from 85 B-lymphoblastoid cell lines homozygous for HLA and peripheral blood granulocytes of 156 Japanese individuals were investigated. Two HLA-B-specific 5′-sided primers (CG4 and CG5) and two 3′-sided primers (CG2 and CG3) were designed for specific amplification of the axon. HLA-B allelss were classified into two groups (groups I and II) according to specific amplification with two types of the 3′-sided primers. The amplified DNAs were hybridized with 23 nonradioactively labeled SSOPs. Based on the hybridization patterns with the SSOPs, 34 HLA-B specificities were divided into 26 epitope combination (EC) groups. Fifteen HLA-B specificities were classified into four EC groups and these HLA-B specificities could not be distinguished from one another in the same EC group. Another 16 HLA-B specificities corresponded one by one to 16 distinct EC groups, and two subtypes of HLA-Bw 75, B27, and Bw48 were also identified enabling the accurate typing of 22 HLA-B alleles at the DNA level. Single-strand conformation polymorphism (SSCP) of the PCR products from group I HLA-B alleles was also investigated. The HLA-B allelss showed distinct electrophoretic patterns in nondenaturing polyacrylamide gels, depending on the nucleotide sequences of the exon 2, indicating that the SSCP analysis may be an alternative, useful and practical HLA-matching system of HLA-B specificity in tissue transplantation.",
author = "Masayuki Yoshida and Akinori Kimura and Fujio Numano and Takehiko Sasazuki",
year = "1992",
month = "1",
day = "1",
doi = "10.1016/0198-8859(92)90025-I",
language = "English",
volume = "34",
pages = "257--266",
journal = "Human Immunology",
issn = "0198-8859",
publisher = "Elsevier Inc.",
number = "4",

}

TY - JOUR

T1 - Polymerase-chain-reaction-based analysis of polymorphism in the HLA-B gene

AU - Yoshida, Masayuki

AU - Kimura, Akinori

AU - Numano, Fujio

AU - Sasazuki, Takehiko

PY - 1992/1/1

Y1 - 1992/1/1

N2 - The polymerase chain reaction (PCR) in combination with the sequence-specific oligonucleotide probe (SSOP) was applied to analyze the polymorphism in the axon 2 of the HLA-B gene. In this study, genomic DNAs from 85 B-lymphoblastoid cell lines homozygous for HLA and peripheral blood granulocytes of 156 Japanese individuals were investigated. Two HLA-B-specific 5′-sided primers (CG4 and CG5) and two 3′-sided primers (CG2 and CG3) were designed for specific amplification of the axon. HLA-B allelss were classified into two groups (groups I and II) according to specific amplification with two types of the 3′-sided primers. The amplified DNAs were hybridized with 23 nonradioactively labeled SSOPs. Based on the hybridization patterns with the SSOPs, 34 HLA-B specificities were divided into 26 epitope combination (EC) groups. Fifteen HLA-B specificities were classified into four EC groups and these HLA-B specificities could not be distinguished from one another in the same EC group. Another 16 HLA-B specificities corresponded one by one to 16 distinct EC groups, and two subtypes of HLA-Bw 75, B27, and Bw48 were also identified enabling the accurate typing of 22 HLA-B alleles at the DNA level. Single-strand conformation polymorphism (SSCP) of the PCR products from group I HLA-B alleles was also investigated. The HLA-B allelss showed distinct electrophoretic patterns in nondenaturing polyacrylamide gels, depending on the nucleotide sequences of the exon 2, indicating that the SSCP analysis may be an alternative, useful and practical HLA-matching system of HLA-B specificity in tissue transplantation.

AB - The polymerase chain reaction (PCR) in combination with the sequence-specific oligonucleotide probe (SSOP) was applied to analyze the polymorphism in the axon 2 of the HLA-B gene. In this study, genomic DNAs from 85 B-lymphoblastoid cell lines homozygous for HLA and peripheral blood granulocytes of 156 Japanese individuals were investigated. Two HLA-B-specific 5′-sided primers (CG4 and CG5) and two 3′-sided primers (CG2 and CG3) were designed for specific amplification of the axon. HLA-B allelss were classified into two groups (groups I and II) according to specific amplification with two types of the 3′-sided primers. The amplified DNAs were hybridized with 23 nonradioactively labeled SSOPs. Based on the hybridization patterns with the SSOPs, 34 HLA-B specificities were divided into 26 epitope combination (EC) groups. Fifteen HLA-B specificities were classified into four EC groups and these HLA-B specificities could not be distinguished from one another in the same EC group. Another 16 HLA-B specificities corresponded one by one to 16 distinct EC groups, and two subtypes of HLA-Bw 75, B27, and Bw48 were also identified enabling the accurate typing of 22 HLA-B alleles at the DNA level. Single-strand conformation polymorphism (SSCP) of the PCR products from group I HLA-B alleles was also investigated. The HLA-B allelss showed distinct electrophoretic patterns in nondenaturing polyacrylamide gels, depending on the nucleotide sequences of the exon 2, indicating that the SSCP analysis may be an alternative, useful and practical HLA-matching system of HLA-B specificity in tissue transplantation.

UR - http://www.scopus.com/inward/record.url?scp=0026467260&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026467260&partnerID=8YFLogxK

U2 - 10.1016/0198-8859(92)90025-I

DO - 10.1016/0198-8859(92)90025-I

M3 - Article

VL - 34

SP - 257

EP - 266

JO - Human Immunology

JF - Human Immunology

SN - 0198-8859

IS - 4

ER -